LOCUS       sequence-487871-        6248 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  pTwist CMV BetaGlobin encoding nivolumab heavy chain with LALA P329G
            knob Fc and HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6248)
  AUTHORS   Stark JC, Gray MA, Wisnovsky S, Ibarlucea-Benitez I, Lustig M, Riley
            NM, Ribi MK, Errington WJ, Bruncsics B, Sarkar CA, Valerius TV, 
            Ravetch JV, Bertozzi CR
  TITLE     Antibody-lectin chimeras for glyco-immune checkpoint blockade
  JOURNAL   bioRxiv 2022.10.26.513931; doi: 
            https://doi.org/10.1101/2022.10.26.513931
REFERENCE   2  (bases 1 to 6248)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2022.10.26.513931; doi: https://doi.org/10.1101/2022.10.26.513931"
COMMENT     Sequence Label: pTCB-nivoHCLALAPGknobHA
FEATURES             Location/Qualifiers
     source          1..6248
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1393..1419
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1472..1593
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1483..1502)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1537..1556
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1708..1754
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1784..1920
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1965..2036
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2262..2850)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2342..2361)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3596..3664,3665..4456)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3814..3833)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4469..4605
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4757..4828
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4833..4879)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4897..4945
                     /note="synthetic polyadenylation signal"
     enhancer        5026..5329
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5331..5533
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5484..5504
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          5667..6142
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
ORIGIN
        1 ctcgagatgg gatggtcatg tatcatcctt tttctagtag caactgcaac cggtgtacat
       61 tcccaagtgc agctcgttga atcaggtggg ggtgtggtac aacctgggag atcactgcga
      121 ctggattgca aggcttccgg tatcacgttt tcaaactcag gtatgcattg ggtccgacag
      181 gcaccaggaa aaggactcga gtgggtagcc gtaatctggt atgacggctc aaaacgctac
      241 tatgcggata gtgtaaaagg aaggttcacg atctcacgcg ataactctaa gaacaccctg
      301 ttccttcaga tgaactccct gcgagcagaa gatacggcag tttattattg tgcaaccaac
      361 gatgattatt ggggccaggg aaccttggtt accgtctcat cagctagcac caagggccca
      421 tcggtcttcc ccctggcacc ctcctccaag agcacctctg ggggcacagc ggccctgggc
      481 tgcctggtca aggactactt ccccgaaccg gtgacggtgt cgtggaactc aggcgccctg
      541 accagcggcg tgcacacctt cccggctgtc ctacagtcct caggactcta ctccctcagc
      601 agcgtggtga ccgtgccctc cagcagcttg ggcacccaga cctacatctg caacgtgaat
      661 cacaagccca gcaacaccaa ggtggacaag aaagttgagc ccaaatcttg tgacaaaact
      721 cacacatgcc caccgtgccc agcacctgaa gcagccgggg gaccgtcagt cttcctcttc
      781 cccccaaaac ccaaggacac cctcatgatc tcccggaccc ctgaggtcac atgcgtggtg
      841 gtggacgtga gccacgaaga ccctgaggtc aagttcaact ggtacgtgga cggcgtggag
      901 gtgcataatg ccaagacaaa gccgcgggag gagcagtaca acagcacgta ccgtgtggtc
      961 agcgtcctca ccgtcctgca ccaggactgg ctgaatggca aggagtacaa gtgcaaggtc
     1021 tccaacaaag ccctcggagc ccccatcgag aaaaccatct ccaaagccaa agggcagccc
     1081 cgagaaccac aggtgtacac cctgccccca tgccgggatg agctgaccaa gaaccaggtc
     1141 agcctgtggt gcctggtcaa aggcttctat cccagcgaca tcgccgtgga gtgggagagc
     1201 aatgggcagc cggagaacaa ctacaagacc acgcctcccg tgctggactc cgacggctcc
     1261 ttcttcctct acagcaagct caccgtggac aagagcaggt ggcagcaggg gaacgtcttc
     1321 tcatgctccg tgatgcatga ggctctgcac aaccactaca cgcagaagag cctctccctg
     1381 tctccgggtg ggtacccata cgatgttcca gattacgctt gagctagctt gactgactga
     1441 gatacagcgt accttcagct cacagacatg ataagataca ttgatgagtt tggacaaacc
     1501 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     1561 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     1621 tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     1681 ggtattggcc catctctatc ggtatcgtag cataacccct tggggcctct aaacgggtct
     1741 tgaggggttt tttgtgcccc tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa
     1801 aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat
     1861 acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa
     1921 ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc aagacattcc tttaatggtc
     1981 ttttctggac accactaggg gtcagaagta gttcatcaaa ctttcttccc tccctaatct
     2041 cattggttac cttgggctat cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga
     2101 tcgacttgtc tgggtttcga ctacgctcag aattgcgtca gtcaagttcg atctggtcct
     2161 tgctattgca cccgttctcc gattacgagt ttcatttaaa tcatgtgagc aaaaggccag
     2221 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2281 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2341 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2401 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2461 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2521 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2581 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2641 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2701 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2761 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2821 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2881 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2941 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3001 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3061 tgtctatttc gttcatccat agttgcattt aaatttccga actctccaag gccctcgtcg
     3121 gaaaatcttc aaacctttcg tccgatccat cttgcaggct acctctcgaa cgaactatcg
     3181 caagtctctt ggccggcctt gcgccttggc tattgcttgg cagcgcctat cgccaggtat
     3241 tactccaatc ccgaatatcc gagatcggga tcacccgaga gaagttcaac ctacatcctc
     3301 aatcccgatc tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg gtgtaccgag
     3361 aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt cagccagtcg
     3421 gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag cctggtcacg
     3481 gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa cgtataatcg
     3541 agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct ttcgcatgag
     3601 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3661 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt
     3721 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3781 acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     3841 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3901 gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3961 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgattggagg
     4021 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4081 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4141 agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc tagcttcccg
     4201 gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4261 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     4321 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     4381 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4441 gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac
     4501 gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact
     4561 tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcc cgaatcgttt
     4621 aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc
     4681 gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc tttttggcag
     4741 cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt ccctctagtg
     4801 gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag
     4861 gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca acacacatcc
     4921 atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc agatctagta
     4981 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     5041 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     5101 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     5161 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     5221 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     5281 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc tgatgcggtt
     5341 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     5401 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     5461 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     5521 tataagcaga gctggtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt
     5581 tgacctccat agaagacacc gggaccgatc cagcctcccc tcgaagctta catgtggtac
     5641 cgagctcgga tcctgagaac ttcagggtga gtctatggga cccttgatgt tttctttccc
     5701 cttcttttct atggttaagt tcatgtcata ggaaggggag aagtaacagg gtacacatat
     5761 tgaccaaatc agggtaattt tgcatttgta attttaaaaa atgctttctt cttttaatat
     5821 acttttttgt ttatcttatt tctaatactt tccctaatct ctttctttca gggcaataat
     5881 gatacaatgt atcatgcctc tttgcaccat tctaaagaat aacagtgata atttctgggt
     5941 taaggcaata gcaatatttc tgcatataaa tatttctgca tataaattgt aactgatgta
     6001 agaggtttca tattgctaat agcagctaca atccagctac cattctgctt ttattttatg
     6061 gttgggataa ggctggatta ttctgagtcc aagctaggcc cttttgctaa tcatgttcat
     6121 acctcttatc ttcctcccac agctcctggg caacgtgctg gtctgtgtgc tggcccatca
     6181 ctttggcaaa ggcggccgct gccaagcttc cgagctctcg aattcaaagg aggtaccgcg
     6241 atatctac
//
